Effect Of Nuclear Factor-kappa Research Articles

Effects of nuclear factor-κB signaling pathway on periodontal ligament stem cells under lipopolysaccharide-induced inflammation

ABSTRACT Lipopolysaccharide (LPS) induces inflammatory stress and apoptosis. This study focused on the effect of nuclear factor kappa B (NF-κB) signaling pathway on proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) after LPS induction and its mechanism. We first isolated hPDLSCs from human tooth root samples in vitro. Then, flow cytometry detected positive expression of cell surface antigens CD146 and STRO-1 and negative expression of CD45, suggesting the hPDLSCs were successfully isolated. LPS significantly induced increased apoptosis and diminished proliferation of hPDLSCs. The NF-κB pathway agonist phorbol 12-myristate 13-acetate (PMA) or p65 overexpression inhibited the proliferation of LPS-treated hPDLSCs and promoted apoptosis. PMA also promoted LPS-induced up-regulation of the expression of inflammatory factors TNF-α and IL-6 and down-regulation of the expression of anti-inflammatory factor IL-10. Additionally, LPS was confirmed to lead to a reduction of alkaline phosphatase (ALP) activity, calcium nodules, and expression of osteogenic markers Runt-related transcription factor 2 (Runx2) and osteopontin. This reduction could be promoted by PMA. Western blotting further indicated that PMA could promote LPS-induced decrease of expression of p65 (cytoplasm), and total cellular proteins IKKα and IKKβ in hPDLSCs, while protein expression of p-IκBα (cytoplasm) and p65 (nucleus), and p-IκBα/IκBα ratio was elevated. By contrast, inhibition of the NF-κB pathway (PDTC) or small-interfering RNA targeting NF-κB/p65 (p65 siRNA) showed the opposite results. In conclusion, activation of NF-κB signaling in LPS-induced inflammatory environment can inhibit the proliferation and osteogenic differentiation of hPDLSCs. This study provides a theory foundation for the clinical treatment of periodontitis.

Synergistic Effect of NF-κB Signaling Pathway Inhibitor and Oncolytic Measles Virus Vaccine Strain against Lung Cancer and Underlying Mechanisms

背景与目的肺癌已成为我国发病率和死亡率居首位的恶性肿瘤。能自我复制、选择性杀伤肿瘤的溶瘤病毒，是治疗恶性肿瘤的有效策略，而其中溶瘤麻疹病毒疫苗株因其良好的溶瘤效果，且对正常细胞无损伤或微损伤，已进入几项临床试验。本研究旨在探讨核转录因子κB（nuclear factor kappa B, NF-κB）信号通路抑制剂与溶瘤麻疹病毒疫苗株协同抗肺癌的作用及机制。方法应用Western blot方法检测MV-Edm感染人肺癌细胞A549和H1299并应用细胞自噬相关的siRNA或者应用NF-кB通路抑制剂PS1145后SQSTM1、p-IκBα、IκBα、PARP及BAX的表达水平，运用流式细胞术分析各组细胞凋亡率的变化，同时采用噻唑蓝[3-(4, 5)-dimethylthiahiazo(-z-y1)-3, 5-di-phenytetrazoliumromide, MTT]法检测各组细胞的存活率。结果Western blot结果显示MV-Edm感染人肺癌细胞A549和H1299后，自噬引起NF-κB通路的激活，进而抑制细胞凋亡。抑制细胞自噬可抑制NF-κB通路的激活，MV-Edm感染后p-IκBα表达水平随着感染时间有不同程度的升高，IκBα的表达水平则降低，NF-κB通路抑制剂PS1145可促进人肺癌细胞A549和H1299凋亡（P < 0.01），并增强其溶瘤效果。结论NF-κB信号通路抑制剂PS1145与溶瘤麻疹病毒疫苗株联合可促进人肺癌细胞A549和H1299凋亡，并增强其溶瘤效果。

The protection of NF-κB inhibition on kidney injury of systemic lupus erythematosus mice may be correlated with lncRNA TUG1.

We aimed to know the effect of nuclear factor-kappa B (NF-κB) inhibition on the kidney injury of systemic lupus erythematosus (SLE) mice. Pristane-induced SLE mice were treated with pyrrolidine dithiocarbamate (PDTC, 50 or 100 mg/kg), a NF-κB inhibitor. Histopathological changes were observed by hematoxylin & eosin, Masson and periodic schiff-methenamine stainings. Long noncoding RNA Taurine upregulated gene 1 (LncRNA TUG1) was measured by real-time reverse transcription PCR, NF-κB p65 expression by western blotting, levels of inflammatory cytokines, antinuclear antibodies (ANA), and antidouble stranded DNA (anti-dsDNA) by enzyme-linked immunosorbent assay, and the deposition of IgG and C3 by immunofluorescence. The kidney of SLE mice exhibited interstitial inflammatory cell infiltration, interstitial fibrous proliferation, glomerular mesangial proliferation, and crescent formation, which was mitigated after PDTC administration. The levels of BUN, Cr, ANA, and anti-dsDNA and the pro-inflammatory factors in SLE mice were increased with obvious deposition of IgG and C3, but they were also reversed by PDTC. Furthermore, the NF-κB p65 expression in the nucleus in the SLE mice was decreased with the up-regulation of TUG1 expression and NF-κB p65 expression in the cytoplasm after PDTC treatment. Correlation analysis revealed the negative correlation between the TUG1 expression and NF-κB p65 in the nucleus in the kidney tissues. NF-κB inhibition with PDTC protected against the kidney injury of pristine-induced SLE mice possibly via up-regulating lncRNA TUG1, and further clinical studies are needed to clarify whether NF-κB inhibition may be a therapeutic modality for the kidney injury of SLE.

The reciprocal interaction of sympathetic nervous system and cAMP-PKA-NF-kB pathway in immune suppression after experimental stroke

BackgroundSympathetic nervous system(SNS) is involved in the mechanism of immune suppression after stroke. Furthermore, as the pro-inflammatory effect of nuclear factor kappa B(NF-kB) is inhibited after stroke, which is regulated by cyclic adenosine monophosphate(cAMP) and proteinkinase A(PKA). The cAMP-PKA-NF-kB pathway might play an important role in noradrenergic-mediated immune dysfunction. AimThe purpose of our research is to analyze how SNS interfere with the immune system after acute stroke and the underlying mechanism of cAMP-PKA-NF-kB pathway in regulating the inflammation. Methods32 healthy male Sprague-Dawley rats were divided into 4 groups equally and randomly (1) Sham operation group; (2) middle cerebral artery occlusion; (MCAO) control group; (3) propranolol MCAO group; (4) isopropylarterenol sham group. 72h later after MCAO or sham operation, tumor necrosis factor-α(TNF-α)and interleukine-10(IL-10) in serum as well as cAMP, PKA and NF-kB in spleen cells were tested. ResultsTNF-α decreased while IL-10 increased in serum after acute ischemia stroke (p<0.05). Meanwhile, the levels of cAMP and PKA in spleen both increased in MCAO model while the expression of NF-kB was inhibited (p<0.05). When propranolol was used to inhibit SNS, all of the results reversed (p<0.05). But the reversed results were still significantly different from the sham operation group (p<0.05). Isopropylarterenol administrated rats appeared the same trend as MCAO group when compared to the sham operation group (p<0.05). However, the differences still existed (p<0.05). ConclusionOn account of the SNS activation after stroke, epinephrine activates the expression of cAMP, which further increases the level of PKA. Therefore, the level of nuclear factor NF-kB is down-regulated. Since the pro-inflammatory effect of NF-kB slacked, the immune system may be inhibited after stroke.

The research of BAY11-1082 on inhibition and inducing apoptosis in pancreatic carcinoma of 5-fluorouracil

Objective To study the influence of BAY11-7082 [nuclear factor-kappa B (NF-κB)inhibitor] on the inhibition and inducing apoptosis in pancreatic carcinoma of 5-fluorouracil (5-Fu).Methods The human pancreatic carcinoma SW1990 cells were deal with 5 μmoL/L BAY11-7082 for 1 h before the cells were exposure to 5-Fu.Western blotting was used to detected the phosphoryltion NF-κB p65.After the cells were exposure to 50,200 μg/L,1.0 mg/L 5-Fu,methyl thiazol tetrazolium (MTT)was used to detect the inhibition rate and real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect the expression mRNA of anti-apoptosis gene B cell lymphoma/leukemia-2 (bcl-2).The level of bcl-2 protein was detected by using Western blotting and flow cytometry was used to detect the apoptosis rate of SW1990.Results After the effect of nuclear factor-kappa B (BAY11-7082),the expression of phosphoryhion NF-κB p65 was decreased obviously.The effect of MTT showed that the inhibition rate of 5-Fu on SW1990 elevated significantly compared with the control groups (P ＜ 0.05).The expression of bcl-2 reduced obviously in protein and mRNA level (P ＜ 0.05).The apoptosis rate the groups that was exposure to BAYll-7082 were (16.79 ± 1.23)％,(26.82 ± 1.26)％,(38.20 ±5.89)％,which elevated significantly compared with control groups (8.64 ± 0.30)％,(14.02 ±0.32)％,(24.13± 1.60)％,(P＜0.05).Conclusion The level of phosphorylation NF-κB p65 reduced after the effect of BAY11-7082,which might increase the sensitivity of SW1990 to 5-Fu and elevate the apoptosis rate of SW1990 after the effect of 5-Fu. Key words: Pancreatic carcinoma; Nuclear factor-kappa B inhibitor; 5-fluorouracil; Apoptosis

Abstract P5-09-04: The effect of nuclear factor kappa B activation on cell proliferation of estrogen receptor positive breast cancer cell lines with different HER2-status

Abstract Introduction:Endocrine treatment is the mainstay of therapy for patients with Estrogen Receptor positive (ER+) breast cancer.Unfortunately, primary and secondary resistance to endocrine therapy are a major clinical problem. Previous studies have suggested a possible role for NFkB transcription factor family members in this resistance due to their effect on ER signaling. In this study, we aim to elucidate the interaction between both transcription factor families (ER and NFkB) and the consequence thereof on the growth pattern of ER+ cell lines. Materials and methods:For this study, 2 ER+ breast cancer cell lines (i.e. MCF7 and MDA-MB-361) were selected based on a documented attenuated NFkB activity in these cell lines and a different ErbB2 status.Prior to treatment with estrogen (E2), TNFα (NFkB activation) and/or parthenolide (NFkB inhibition), cells were seeded in 48 well plates. Proliferation was measured by means of a crystal violet assay. ELISA experiments to measure NFkB DNA-binding in nuclear protein extracts and qRT-PCR for NFkB target genes was performed. Results:Treatment of MCF7cells with different concentrations of TNFα initially increased cell proliferation, but after 48hrs, apoptosis occurred. Evaluation of NFkB DNA-binding showed a clear increase in RelA (p65), RelB (p62) and NFkB1 (p50) DNA-binding after 4hrs and up until but no longer than 24hrs of treatment. Results were confirmed by qRT-PCR for NFkB target genes.Due to the dynamics of NFkB activation secondary to TNFα treatment, we limited further experiments to 24hrs of treatment. Combination of E2and TNFα gave an additive effect in MCF7 cell proliferation compared to E2 alone, an effect counteracted by parthenolide in a concentration dependent manner. In the ErbB2+ cell line MDA-MB-361 our initial results suggest that addition of TNFα does not affect E2-driven cell proliferation. Conclusion: Our results so far suggest a different, ErbB2-dependent role for NFkB on the proliferation of ER+ breast cancer cells. Interestingly, the effect of NFkB seems to depend on both RelA- and RelB-dependent pathways. Experiments are currently being performed to elaborate on the present results. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-09-04.

Effect of inhibitory nuclear factor kappa B on apoptosis of prostate cancer cell line PC-3M inducted by TRAIL

Objective To investigate the suppressive effect of pyrrolidine dithiocarbamate on nuclear factor kappa B and to evaluate the effect of enhancement of PDTC on the apoptosis of androgen-independent prostate cancer cell line PC-3M when tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is used. Methods PC-3M cells were divided to different groups according to the treatment with TRAIL (25,50,100 μg/L) or PDTC (25,50,100 μmol/L) or both of them [25/25,25/50,25/100,50/ 25,50/50,50/100 PDTC( μmol/L) ,TRAIL( μg/L) ]. The nuclear translocation of nuclear factor kappa B was evaluated by cell immunohistochemical method and electrophoretic mobility shift assay ( EMSA). The analys of apoptosis was carried out by MTT test and flow cytometry. Results EMSA and cell immunohistochemical analysis showed that the translocation of nuclear factor kappa B can be greatly activated when PC3M cells were treated with TRAIL. The pretreatment of PDTC can block the nuclear translocation and eliminate the protection to cancer cells in apoptosis. In the meantime, as a stong suppressor of nuclear factor kappa B, PDTC is less toxic. The killing effect of TRAIL + PDTC group is obviously more powerful than TRAIL-single group (P <0. 01) . Conclusion The effect of TRAIL will be remarkably neutralized by the activation of nuclear factor kappa B when it deal with androgen-independent prostate cancer cell. On the other hand,TRAIL can significantly and efficiently induce the apoptosis of PC-3M cells when PDTC is pretreated to inhibit the activation of nuclear factor kappa B. Key words: Prostate neoplasm; Carcinoma; Tumor necrosis factor; Nuclear factor kappa B

Effect of nuclear factor-kappa B decoy oligodeoxynncleotides on respiratory function and cytokine expression after severe lung contusion in rabbits

Objective To explore the effect of nuclear factor-kappa B （ NF-κB ） decoy oligodeoxynueleotides （ODN） on respiratory function and expressions of IL-1β and IL-13 in serum following severe lung contusion in rabbits. Methods A total of 40 New-Zealand rabbits were randomly divided into four groups, ie, severe lung contusion group （Group A, n = 12）, severe lung contusion with NF-κB scrambled decoy ODN intervention group （ Group B, n = 12） , severe lung contusion with sense NF- B decoy ODN intervention group （Group C, n = 12） and nomlal control group （Group D, n =4）. After the contusion model was set up, the sense and scrambled NF-κB decoy ODN were infused into the rabbits via the jugular veins in different groups, with 20 g per experimental rabbit. After contusion, respiratory frequency, tidal volume, airway pressure, respiration flow rate curve and end expiration nitric oxide concentration were detected at 1, 2, 3 and 4 hours. The expressions of IL-1β and IL-13 in serum were observed by means of ELISA. Results After sense NF-κB decoy ODN intervention, alveolar ventilation, arterial PO2 and pulmonary compliance were improved, compared with Group A and Group B, with statistical difference （ P 〈 0. 01 ）. The expression of IL-1β was decreased and that of IL-13 increased after sense NF-κB decoy ODN intervention to the severe lung contusion, compared with Groups A and B, with statistical difference （P 〈0.01 ）. The expression of IL-1β was increased to peak level at 1 hour after contusion, which continued to the end of the experiment. While expression of IL-13 was decreased at 1 hour after contusion and reached the minimum level at 4 hours. With intervention with sense decoy ODN, the increased expression of IL-1β was down-regulated, but expression of IL-13 remained at high level, with statistical difference compared with Group A and Group B （ P 〈 0. 01 ）. Conclusions Intervention with sense NF-κB decoy ODN can significantly protect the respiratory function, reduce the expression of IL-1β, and increase expression of IL-13 after severe lung contusion. Key words: Thoracic injuries ; Nuclear factor-kappa B ; Decoy oligodeoxynucleotides ; Respiratory mechanics; Pulmonary ventilation; Pulmonary gas exchange; Inflammation mediators

Effect of nuclear factor kappa B on intercellular adhesion molecule-1 expression and neutrophil infiltration in lung injury induced by intestinal ischemia/reperfusion in rats

To investigate the role of nuclear factor kappa B (NF-kappaB) in the pathogenesis of lung injury induced by intestinal ischemia/reperfusion (I/R), and its effect on intercellular adhesion molecule-1 (ICAM-1) expression and neutrophil infiltration. Twenty-four Wistar rats were divided randomly into control, I/R and pyrrolidine dithiocarbamate (PDTC) treatment groups, n=8 in each. I/R group and PDTC treatment group received superior mysenteric artery (SMA) occluding for 1 h and reperfusion for 2 h. PDTC group was administrated with intraperitoneal injection of 2% 100 mg/kg PDTC 1 h before surgery. Lung histology and bronchia alveolus lung fluid (BALF) protein were assayed. Serum IL-6, lung malondialdehyde (MDA) and myeloperoxidase (MPO) as well as the expression level of NF-kappaB and ICAM-1 were measured. Lung injury induced by intestinal I/R, was characterized by edema, hemorrhage and neutrophil infiltration as well as by the significant rising of BALF protein. Compared to control group, the levels of serum IL-6 and lung MDA and MPO increased significantly in I/R group (P=0.001). Strong positive expression of NF-kappaB p65 and ICAM-1 was observed. After the administration of PDTC, the level of serum IL-6, lung MDA and MPO as well as NF-kappaB and ICAM-1 decreased significantly (P<0.05) when compared to I/R group. The activation of NF-kappaB plays an important role in the pathogenesis of lung injury induced by intestinal I/R through upregulating the neutrophil infiltration and lung ICAM-1 expression. PDTC as an inhibitor of NF-kappaB can prevent lung injury induced by intestinal I/R through inhibiting the activity of NF-kappaB.

Effect of nuclear factor-kappa B on cell survival of SOD1 TG astrocytes after oxygen glucose deprivation

Effect of nuclear factor-kappa B on cell survival of SOD1 TG astrocytes after oxygen glucose deprivation

Effect of nuclear factor-kappa B inhibitors and peroxisome proliferator-activated receptor-gamma ligands on PTHrP release from human fetal membranes

Parathyroid hormone-related protein (PTHrP) has been implicated in many processes during normal and pathological pregnancies. In the human fetal membranes, PTHrP exhibits cytokine-like actions. We have recently shown that inhibitors of the nuclear factor-kappa B (NF-κB) and activators of the peroxisome proliferator-activated receptor (PPAR)-γ signalling pathways down-regulate cytokine release from human gestational tissues. Therefore, the aim of this study was to determine whether NF-κB and PPAR-γ also regulate PTHrP release from human fetal membranes. Human amnion and choriodecidua explants were incubated in the absence (control) or presence of two known NF-κB inhibitors (1, 5 and 10 mM sulphasalazine (SASP) or 5, 10 and 15 mM N-acetyl-cysteine (NAC)), and two PPAR-γ ligands (15 and 30 μM 15-deoxy-Δ 12,14-PGJ 2 (15d-PGJ 2) or 15 and 30 μM troglitazone), under basal conditions. After 18 h incubation, the tissues were collected and NF-κB p65 DNA binding activity in nuclear extracts was assessed by ELISA, and the incubation medium was collected and the release of PTHrP was quantified by RIA. Treatment of amnion and choriodecidual tissues with SASP concentrations greater than 5 mM, 15 mM NAC, 30 μM 15d-PGJ 2 and 30 μM troglitazone significantly reduced the release of PTHrP ( p<0.05). This study demonstrates that PTHrP release from human fetal membranes is regulated by inhibitors of NF-κB, and ligands of PPAR-γ.

Effect of nuclear factor-kappa B on vascular endothelial growth factor mRNA expression of human pulmonary artery smooth muscle cells in hypoxia.

In order to investigate the effect of nuclear factor-kappa B (NF-kappaB) on vascular endothelial growth factor (VEGF) mRNA expression of human pulmonary artery smooth muscle cells (HPASMCs) in hypoxia, the cultured HPASMCs in vitro were stimulated with pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB. The NF-kappaB p65 nuclei positive expression was detected by immunocytochemical technique. The IkappaBalpha protein expression was measured by Western blot. RT-PCR was used to detect the VEGF mRNA expression of HPASMCs. The results showed that no significant change was observed in the NF-kappaB p65 nuclei positive expression of cultured HPASMCs during 6 h-24 h in normoxia, but the levels of NF-kappaB p65 nuclei positive expression of cultured HPASMCs were significantly increased in hypoxia groups as compared with those in all normoxia groups (P<0.05). The IkappaBalpha protein expression of cultured HPASMCs showed no significant change during 6 h-24 h in normoxia, but significantly decreased in hypoxia as comapred with that in normoxia groups (P<0.05). PDTC (1 to 100 micromol/L) could inhibit the VEGF mRNA expression of HPASMCs in a concentration-dependent manner in hypoxia. In conclusion, NF-kappaB can be partly translocation activated from cytoplasm into nuclei in the cultured HPASMCs under hypoxia. The inhibition of NF-kappaB activation can decrease the VEGF mRNA expression. It is suggested that the activation of NF-kappaB is involved in the VEGF mRNA expression of HPASMCs under hypoxia.